Protease, Staph aureus (Endoproteinase Glu-C) Assay

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Protease, Staph aureus (Endoproteinase Glu-C) Assay

Method

Enzyme activity is determined by the casein digestion assay described by Drapeau (1976). Since substrate grade caseins can vary from lot to lot and according to the manner produced, the standardization of casein digestion assays has been difficult to achieve. The use of a reference enzyme preparation is suggested. One unit is that amount of enzyme which releases acid soluble fragments equivalent to 0.001 A₂₈₀ per minute at 37°C and pH 7.8 under the specified conditions.

Reagents

1% Casein in 0.05 M Tris⋅PO₄ buffer, pH 7.8. (Dissolve 1 gram "Hammersten" casein in 50 ml 0.01 N NaOH with gentle heating and stirring. Add 40 ml reagent grade water and 5.0 ml 1.0 M Tris. Adjust pH to 7.8 with H₃PO₄ and q.s. to 100 ml.)
10% Trichloroacetic acid (TCA)

Enzyme

Dissolve at 1 mg/ml in reagent grade water.

Procedure

Equilibrate a series of tubes with 5.0 ml of 1% casein at 37°C for 5 minutes. At zero time add 10 ul or 20 ul of enzyme. Mix. Include a reagent blank. Exactly ten minutes after adding sample, stop reaction by adding 5.0 ml TCA. Mix. Allow tubes to stand ten minutes and then filter. Read A₂₈₀ of the filtrate.

Calculation

\frac{\text{Units}}{\text{mg}}\text{ = }\frac{\underset{\text{Test}}{\text{(A}_{280}}\text{ - }\underset{\text{Reagent Blank}}{\text{A}_{280}\text{)}}}{\text{10 min x mg enzyme in reaction}}

Protease, Staph aureus (Endoproteinase Glu-C) Products

Description

Protease, S. aureus, Sequencing Grade

Source:

Staphylococcus aureus V8

Chromatographically purified according to Drapeau, G, et.al, JBC, 247, 6720 (1972). Supplied as a lyophilized powder.

Store at 2-8°C.

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Activity

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Certificate of Analysis

Code

STSEQ

Product details

LS02126

5x10 ug

$192.00

LS02128

5x50 ug

$520.00

LS02129

Bulk

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Description

Protease, S. Aureus (Endoproteinase Glu-C)

Source:

Staphylococcus aureus V8

Chromatographically purified according to Drapeau, G., Boily, Y., and Houmard, J.: JBC, 247, 6720 (1972). A lyophilized powder.

Store at 2-8°C.

…

Activity

≥500 units per mg dry weight

Certificate of Analysis

Code

STAP

Product details

LS003608

1 mg

$76.00

LS003605

5 mg

$270.00

LS003606

Bulk

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Protease, Staph aureus (Endoproteinase Glu-C) - Assay

Protease, Staph aureus (Endoproteinase Glu-C) Assay

Protease, Staph aureus (Endoproteinase Glu-C) Products